Jinn Tzyy-Rong, Kao Suey-Sheng, Tseng Yin-Chin, Chen Ying-Ju, Wu Tzong-Yuan
Graduate Institute of Chinese Medical Science, China Medical University, Taichung, Taiwan.
Biotechnol Prog. 2009 Mar-Apr;25(2):384-9. doi: 10.1002/btpr.148.
The baculovirus-insect expression system is a popular tool for recombinant protein production. The standard method for infecting insect larvae with recombinant baculovirus for protein production involves either feeding occlusion bodies or injecting budded virus into the cuticle. In this study, we showed that the recombinant Autographa californica multiple nucleopolyhedrovirus (AcMNPV) at titers >10(8) pfu/mL efficiently infected Trichoplusia ni (T. ni) larvae through aerosol inoculation of budded virus at a pressure of 5.5 x 10(4) Pa. The dipping T. ni larvae in virus-containing solution efficiently infected them. These results indicate that surface contamination, either by aerosol or dipping, lead to infection via spiracles. The aerosol infection route for AcMNPV was restricted to T. ni and Plutella xylostella larvae, whereas Spodoptera litura and Helicoverpa armigera larvae were resistant to this inoculation process. The yields of the reporter proteins DsRed and EGFP from T. ni larvae following aerosol infection were nearly identical to those following oral feeding or injection. This alternative baculovirus infection strategy facilitates recombinant protein and virus production by insect larvae.
杆状病毒-昆虫表达系统是一种用于生产重组蛋白的常用工具。用重组杆状病毒感染昆虫幼虫以生产蛋白的标准方法包括喂食包涵体或将出芽病毒注射到表皮中。在本研究中，我们表明，滴度>10(8) pfu/mL的重组苜蓿银纹夜蛾多核多角体病毒(AcMNPV)在5.5 x 10(4) Pa的压力下通过气溶胶接种出芽病毒有效地感染了粉纹夜蛾(T. ni)幼虫。将粉纹夜蛾幼虫浸入含病毒溶液中能有效地感染它们。这些结果表明，通过气溶胶或浸入造成的表面污染会导致通过气门感染。AcMNPV的气溶胶感染途径仅限于粉纹夜蛾和小菜蛾幼虫，而斜纹夜蛾和棉铃虫幼虫对这种接种过程具有抗性。气溶胶感染后粉纹夜蛾幼虫中报告蛋白DsRed和EGFP的产量与口服喂食或注射后的产量几乎相同。这种替代性的杆状病毒感染策略有助于昆虫幼虫生产重组蛋白和病毒。
